Summary
The analysis of the expression of virulence genes and the elucidation of metabolic and regulatory pathways of Staphylococcus aureus provide us with important information about the interaction between the pathogen and its host, mechanisms by which this organism causes diseases, and the resistance to antibiotics. In order to investigate regulatory networks of S. aureus, we analyze the cytoplasmic and extracellular proteome by using two-dimensional (2D) gel analyses combined with matrix-assisted laser ionization–time-of-flight mass spectrometry (MALDI–TOF MS). Gel-based proteomics is an extremely valuable tool in microbial physiology that can, in combination with various visualization and quantitation software packages, very rapidly provide comparative and quantitative data for multi-sample comparison.
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References
Kuroda, M., Ohta, T., Uchiyama, I., Baba, T., Yuzawa, H., Kobayashi, I., Cui, L., Oguchi, A., Aoki, K., Nagai, Y., Lian, J., Ito, T., Kanamori, M., Matsumaru, H., Maruyama, A., Murakami, H., Hosoyama, A., Mizutani-Ui, Y., Takahashi, N. K., Sawano, T., Inoue, R., Kaito, C., Sekimizu, K., Hirakawa, H., Kuhara, S., Goto, S., Yabuzaki, J., Kanehisa, M., Yamashita, A., Oshima, K., Furuya, K., Yoshino, C., Shiba, T., Hattori, M., Ogasawara, N., Hayashi, H., and Hiramatsu, K. (2001) Whole genome sequencing of meticillin-resistant Staphylococcus aureus. Lancet 357, 1225–1240.
O’Farrell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem.250, 4007–4021.
Klose, J. (1975) Protein mapping by combined isoelectric focusing and electrophoresis of mouse tissues. A novel approach to testing for induced point mutations in mammals. Humangenetik 26, 231–243.
Kohler, C., Wolff, S., Albrecht, D., Fuchs, S., Becher, D., Büttner, K., Engelmann, S., and Hecker, M. (2005) Proteome analyses of Staphylococcus aureus in growing and non-growing cells: a physiological approach. Int. J. Med. Microbiol.295, 547–565.
Hecker, M. and Engelmann, S. (2006) Physiological proteomics of Bacillus subtilis and Staphylococcus aureus: towards a comprehensive understanding of cell physiology and pathogenicity, in Pathogenomics (Hacker, J. and Dobrindt, U., eds.), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, pp. 43–68.
Ziebandt, A. K., Becher, D., Ohlsen, K., Hacker, J., Hecker, M., and Engelmann, S. (2004) The influence of agr and sigma(B) in growth phase dependent regulation of virulence factors in Staphylococcus aureus. Proteomics 4, 3034–3047.
Novick, R. P. (2003) Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol. Microbiol.48, 1429–1449.
Ziebandt, A. K., Weber, H., Rudolph, J., Schmid, R., Höper, D., Engelmann, S., and Hecker, M. (2001) Extracellular proteins of Staphylococcus aureus and the role of SarA and sigma B. Proteomics 1, 480–493.
Rogasch, K., Röhmling, V., Pané-Farré, J., Höper, D., Weinberg, C., Fuchs, S., Schmudde, M., Bröker, B. M., Wolz, C., Hecker, M., and Engelmann, S. (2006) The influence of the two component system SaeRS on global gene expression in two different Staphylococcus aureus strains. J. Bacteriol. 188, 7742–7758.
Sibbald, M. J. J. B., Ziebandt, A. K., Engelmann, S., Hecker, M., de Jong, A., Harmsen, H. J. M., Raangs, G. C., Stokroos, I., Arends, J. P., Dubois, J. Y. F., and van Dijl, J. M. (2006) Mapping of pathways to staphylococcal pathogenesis by comparative secretomics. Microbiol. Mol. Biol. Rev.70, 755–788.
Gertz, S., Engelmann, S., Schmid, R., Ohlsen, K., Hacker, J., and Hecker M. (1999) Regulation of sigmaB-dependent transcription of sigB and asp23 in two different Staphylococcus aureus strains. Mol. Gen. Genet.261, 558–566.
Candiano, G., Bruschi, M., Musante, L., Santucci, L., Ghiggeri, G. M., Carnemolla, B., Orecchia, P., Zardi, L., and Rigetti, P. G. (2004) Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis. Electrophoresis 25, 1327–1333.
Blum, H., Beier, H., and Gross, H. J. (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8, 93–99.
Eymann, C., Dreisbach, A., Albrecht, D., Bernhardt, J., Becher, D., Gentner, S., Tam, L. T., Böttner, K., Buurman, G., Scharf, C., Venz, S., Völker, U., and Hecker, M. (2004) A comprehensive proteome map of growing Bacillus subtilis cells. Proteomics 4, 2849–2876.
Bernhardt, J., Böttner, K., Scharf, C., and Hecker, M. (1999) Dual channel imaging of two-dimensional electropherograms in Bacillus subtilis. Electrophoresis 20, 2225–2240.
Acknowledgments
We are very grateful to Christian Kohler, Anne-Kathrin Ziebandt, Harald Kusch, Stephan Fuchs, and Robert S. Jack for critical reading the manuscript and preparing of figures. We thank all co-workers and students for their excellent work on proteome analyses. Furthermore, we thank the Decodon GmbH for providing the Delta2D software. This work was supported by grants from the Bildungsministerium für Bildung und Forschung (031U107A/-207A; 031U213B), the DFG (GK212/3-00), the Land Mecklenburg-Vorpommern, and the Fonds der Chemischen Industrie to M.H. and S.E.
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Engelmann, S., Hecker, M. (2008). Proteomic Analysis to Investigate Regulatory Networks in Staphylococcus aureus . In: DeLeo, F.R., Otto, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology™, vol 431. Humana Press. https://doi.org/10.1007/978-1-60327-032-8_3
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DOI: https://doi.org/10.1007/978-1-60327-032-8_3
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